*2.5. Bioinformatic Analysis*

Sequencing data were subjected to NonPareil 3 [23] for sequencing depth assessment and later processed with Qiime2 (version 2018.11) package [24]. The reads were imported into Qiime2 and run through the dada2 plugin to obtain amplicon sequence variants (ASV) [25]. Taxonomy was assigned for each of the ASVs using a pre-trained naive Bayes classifier, based on the Silva 132 99% database [26], which was trimmed to include only the V3 and V4 regions of the 16S rRNA gene, bound by the Bakt\_341F and Bakt\_805R primer sequences. Alfa and beta diversity metrics were generated using the following Qiime2 plugins: phylogeny (including mafft aligner and FastTree tool), diversity and emperor [27–29].
